Back to Search
Start Over
Impact of whole grain highland hull-less barley on the denaturing gradient gel electrophoresis profiles of gut microbial communities in rats fed high-fat diets.
- Source :
-
Microbiology spectrum [Microbiol Spectr] 2024 Jun 04; Vol. 12 (6), pp. e0408923. Date of Electronic Publication: 2024 May 15. - Publication Year :
- 2024
-
Abstract
- Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) is a traditional non-culture technique that can provide a fingerprint of the microbial community. In the field of gut microbiota analysis, PCR-DGGE still holds potential for development. In the present study, we utilized an improved nested PCR-DGGE approach targeting the V3 region of 16S ribosomal DNA to investigate the impact of whole grain highland hull-less barley (WHLB), a cereal known for its significant hypocholesterolemic effect, on the gut microbiota profiles of high-fat diet rats. Seventy-two male Sprague-Dawley rats were divided into four groups and fed a normal control diet, a high-fat diet, or a high-fat diet supplemented with a low or high dose of WHLB for 4 or 8 weeks. The results revealed that the dominant bands varied among different dose groups and further changed with different treatment times. The compositions of bacterial communities in feces and cecal content were similar, but the dominant bacterial bands differed. After performing double DGGE, extracting the bands, sequencing the DNA, and aligning the sequences, a total of 19 bands were classified under the Firmicutes and Bacteroidetes phyla, while two bands were identified as unclassified uncultured bacteria. The relative abundance of Lactobacillus gasseri , Uncultured Prevotella sp., and Clostridium sp. increased following the administration of WHLB. Illumina-based sequencing was employed to assess the reliability of DGGE, demonstrating its reliability in analyzing the dominant taxonomic composition, although it may have limitations in accurately detecting the alpha diversity of bacterial species.<br />Importance: While next-generation sequencing has overshadowed polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), the latter still holds promise for advancing gut microbiota analysis due to its unique advantages. In this study, we used optimized nested PCR-DGGE to investigate the gut microbiota profile of high-fat diet rats after administering whole grain highland hull-less barley. High-throughput sequencing was employed to validate the DGGE results. Our results proved the reliability of PCR-DGGE for analyzing the dominant taxonomic composition while also providing visual evidence of a notable relationship between the composition of cecal and fecal microbial communities, highlighting substantial differences in both richness and abundance.<br />Competing Interests: The authors declare no conflict of interest.
- Subjects :
- Animals
Male
Rats
Feces microbiology
DNA, Bacterial genetics
Polymerase Chain Reaction methods
Cecum microbiology
Hordeum microbiology
Rats, Sprague-Dawley
Gastrointestinal Microbiome genetics
Diet, High-Fat
Bacteria classification
Bacteria genetics
Bacteria isolation & purification
Denaturing Gradient Gel Electrophoresis
RNA, Ribosomal, 16S genetics
Whole Grains
Subjects
Details
- Language :
- English
- ISSN :
- 2165-0497
- Volume :
- 12
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Microbiology spectrum
- Publication Type :
- Academic Journal
- Accession number :
- 38747621
- Full Text :
- https://doi.org/10.1128/spectrum.04089-23